| Project/Area Number |
09670226
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Experimental pathology
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| Research Institution | KOBE UNIVERSITY |
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Principal Investigator |
KITAZAWA Sohei KOBE UNIVERSITY SCHOOL OF MEDICINE,2ND DEPARTMENT OF PATHOLOGY,ASSISTANT PROFESSOR, 医学部, 講師 (90186239)
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| Project Period (FY) |
1997 – 1998
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| Project Status |
Completed (Fiscal Year 1998)
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| Budget Amount *help |
¥3,200,000 (Direct Cost: ¥3,200,000)
Fiscal Year 1998: ¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 1997: ¥2,600,000 (Direct Cost: ¥2,600,000)
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| Keywords | promoter / gene expression / cyclin D1 / CpG island / サイクリンD_1 / CpG Island / メチル化シトシン / レポータ遺伝子 / 遺伝子導入 / ラット / 遺伝子クローニング |
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| Research Abstract |
We have shown precise localization of methylated cytosine in the 5'-flanking region of rat cyclin D1 gene by molecular pathological technique. In a series of chemical carcinogen-induced rat leukemic cell lines established in our laboratory, we found that the expression of the cyclin D1 integrin varied among cell lines tested. We found no gene amplification and rearrangement in those cell lines. We therefore cloned 5'-flanking region containing cyclin Dl gene promoter to elucidate the mechanism of overexpression. When we sequenced rat genomic DNA,. we found that cyclin Dl gene promoter contained two continuous Spl sites with numerous CpG loci. We therefore speculated that CpG methylation might play an important role in the regulation of the cyclin D1 gene expression at steady-state level, First we screened the methylation status by restriction enzymatic digestion followed by Southern blot analysis. As we expected, we found that heavy CpG methylation resulted in the suppression of the cyclin D1 gene expression at the steady-state level. Furthermore, demethylation of the CpG loci especially around the two Spl sites up-regulated the steady-state expression of cyclin Dl gene.
We have presented our data at various international congress as well as domestic meetings, and publish a lot of scientific papers for academic journals.
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